Hirono-Hara Yoko, Yui Miyuu, Hara Kiyotaka Y
Department of Environmental and Life Sciences, School of Food and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan.
Graduate Division of Nutritional and Environmental Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan.
AMB Express. 2021 Jan 7;11(1):13. doi: 10.1186/s13568-020-01176-3.
Transglutaminase (TG) catalyzes the formation of cross-links between proteins. TG from Streptoverticillium mobaraense (SmTG) is used widely in food, cosmetic, biomaterial and medical industries. SmTG is occasionally supplied as a mixture with the activator peptide glutathione. Currently, glutathione is industrially produced using a budding yeast, Saccharomyces cerevisiae, because of its intracellular high content of glutathione. In this study, active SmTG was produced together with glutathione in S. cerevisiae. SmTG extracted from S. cerevisiae expressing SmTG showed cross-linking activity when BSA and sodium caseinate were substrates. The cross-linking activity of SmTG increased proportionally as the concentration of added glutathione increased. Furthermore, SmTG was prepared by extracting SmTG from an engineered S. cerevisiae whose glutathione synthetic pathway was enhanced. The SmTG solution showed higher activity when compared with a SmTG solution prepared from a S. cerevisiae strain without enhanced glutathione production. This result indicates that a high content of intracellular glutathione further enhances active SmTG production in S. cerevisiae. S. cerevisiae co-producing SmTG and a higher content of glutathione has the potential to supply a ready-to-use industrial active TG solution.
转谷氨酰胺酶（TG）催化蛋白质之间形成交联。茂原链霉菌转谷氨酰胺酶（SmTG）在食品、化妆品、生物材料和医疗行业中被广泛使用。SmTG偶尔会与激活肽谷胱甘肽混合供应。目前，由于其细胞内谷胱甘肽含量高，谷胱甘肽是利用出芽酵母酿酒酵母进行工业生产的。在本研究中，活性SmTG与谷胱甘肽在酿酒酵母中共同产生。从表达SmTG的酿酒酵母中提取的SmTG以牛血清白蛋白（BSA）和酪蛋白酸钠为底物时表现出交联活性。随着添加的谷胱甘肽浓度增加，SmTG的交联活性成比例增加。此外，通过从谷胱甘肽合成途径增强的工程酿酒酵母中提取SmTG来制备SmTG。与从谷胱甘肽产量未增强的酿酒酵母菌株制备的SmTG溶液相比，该SmTG溶液表现出更高的活性。这一结果表明，细胞内高含量的谷胱甘肽进一步增强了酿酒酵母中活性SmTG的产生。共同产生SmTG和更高含量谷胱甘肽的酿酒酵母有潜力提供即用型工业活性TG溶液。
